Side-specific endothelial-dependent regulation of aortic valve calcification: interplay of hemodynamics and nitric oxide signaling

Am J Pathol. 2013 May;182(5):1922-31. doi: 10.1016/j.ajpath.2013.01.037. Epub 2013 Mar 13.


Arterial endothelial cells maintain vascular homeostasis and vessel tone in part through the secretion of nitric oxide (NO). In this study, we determined how aortic valve endothelial cells (VEC) regulate aortic valve interstitial cell (VIC) phenotype and matrix calcification through NO. Using an anchored in vitro collagen hydrogel culture system, we demonstrate that three-dimensionally cultured porcine VIC do not calcify in osteogenic medium unless under mechanical stress. Co-culture with porcine VEC, however, significantly attenuated VIC calcification through inhibition of myofibroblastic activation, osteogenic differentiation, and calcium deposition. Incubation with the NO donor DETA-NO inhibited VIC osteogenic differentiation and matrix calcification, whereas incubation with the NO blocker l-NAME augmented calcification even in 3D VIC-VEC co-culture. Aortic VEC, but not VIC, expressed endothelial NO synthase (eNOS) in both porcine and human valves, which was reduced in osteogenic medium. eNOS expression was reduced in calcified human aortic valves in a side-specific manner. Porcine leaflets exposed to the soluble guanylyl cyclase inhibitor ODQ increased osteocalcin and α-smooth muscle actin expression. Finally, side-specific shear stress applied to porcine aortic valve leaflet endothelial surfaces increased cGMP production in VEC. Valve endothelial-derived NO is a natural inhibitor of the early phases of valve calcification and therefore may be an important regulator of valve homeostasis and pathology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Aortic Valve / enzymology
  • Aortic Valve / pathology*
  • Aortic Valve / physiopathology
  • Aortic Valve Stenosis / enzymology
  • Aortic Valve Stenosis / pathology*
  • Aortic Valve Stenosis / physiopathology*
  • Calcinosis / enzymology
  • Calcinosis / pathology*
  • Calcinosis / physiopathology*
  • Cell Differentiation
  • Endothelial Cells / pathology*
  • Gels
  • Heart Valves / enzymology
  • Heart Valves / pathology
  • Hemodynamics*
  • Humans
  • Immunohistochemistry
  • Myofibroblasts / metabolism
  • Myofibroblasts / pathology
  • Nitric Oxide / metabolism*
  • Nitric Oxide Synthase Type III / metabolism
  • Osteoblasts / metabolism
  • Osteoblasts / pathology
  • Real-Time Polymerase Chain Reaction
  • Signal Transduction*
  • Staining and Labeling
  • Sus scrofa


  • Gels
  • Nitric Oxide
  • Nitric Oxide Synthase Type III

Supplementary concepts

  • Aortic Valve, Calcification of